Use of Antisense Oligonucleotides: Advantages, Controls, and Cardiovascular Tissue

Abstract

Antisense oligonucleotides are short pieces of synthetic, chemically modified DNA or RNA that are designed to interact by Watson-Crick base pairing with mRNA encoding a targeted protein. During the past 20 years the technology associated with the development of antisense has improved dramatically, and emerging chemistries have made antisense oligonucleotides into powerful and versatile tools to study the function of proteins in living cells. The dramatic increase in novel genomic sequence information that has recently become available has generated enormous opportunities for the development of antisense oligonucleotides capable of altering the expression level of virtually any gene. With this will come a nearly equal opportunity to determine the role of individual proteins in a vast array of cardiovascular disease. The great specificity that these compounds exhibit in vitro suggests that they may also have an exciting future for development into therapeutics useful for the treatment of human disease. This review highlights some of the advances made in the field of antisense research, placing an emphasis on uses and proper controls.

Regulation of rat pial arteriolar smooth muscle relaxation in vivo through multidrug resistance protein 5-mediated cGMP efflux

“…When designing the MRP5 antisense ODN, care was taken to avoid four contiguous G residues, which may impart a secondary structure that can prevent hybridization. Additionally, the ODN was modified with end-phosphorothioation (i.e., 3 bases at the 5Ј end and 3 bases at the 3Ј end), which protects the oligonucleotide from rapid nuclease-mediated destruction, while preserving its ability to promote RNase-H-mediated cleavage of the target mRNA (9). A BLAST analysis was also performed to confirm the specificity of the sequence.…”

Potassium Channels in the Peripheral Microcirculation

“…The use of genomic [52] , [94] and proteomic approaches, applied specifically to the microcirculation, appear to be efficient approachs to examine the large number and the diversity of expression patterns of ion channels in microvascular smooth muscle cells, pericytes, and endothelial cells. In addition, the use of gene silencing methods, such as antisense oligonucleotides [51] and siRNA [108] , in conjunction with conventional pharmacology provides a powerful set of tools to examine the function of specific ion channels in the microcirculation. Indeed, the antisense approach has already proven successful in identification of a component of stretch‐activated cation channels in resistance artery smooth muscle cells in vitro [153] .…”

Cantharidin-induced mitotic arrest is associated with the formation of aberrant mitotic spindles and lagging chromosomes resulting, in part, from the suppression of PP2Aα

“…4. The cationic lipids (DOTMA/DOPE; Lipofectin) were used to facilitate the uptake of the oligonucleotides (24,27,28), and the reduction in PP2A mRNA levels observed in response to treatment was varied. Treatment with some oligonucleotides had little or no effect on the inhibition of PP2A mRNA levels, whereas others had a moderate effect, and a few had pronounced effects (Fig.…”